Myenteric Plexus Of Rat Colon Research Articles

An efficient approach for wholemount preparation of the myenteric plexus of rat colon

BackgroundThe nerve plexus of the enteric nervous system (ENS) plays a crucial part in regulating gastrointestinal functions, such as muscle contractile activity and nutrient absorption. Studying the nerve plexus can provide vital information for research on ENS disorders. Whole-mount preparation is an important technique for investigating the nerve plexus. However, currently available methods are time consuming and highly technical. New methodThis study describes a simple and rapid method for preparing whole mounts of the longitudinal muscle and myenteric plexuses (LMMPs) of rat colon. Integral LMMPs can be easily separated from the underlying layer by using glass rods and wet cotton swabs. ResultsThe proposed method allows the easy separation of the LMMPs in intact sheets. Immunofluorescence histochemical staining of whole mounts enable clear visualization of enteric ganglia, nerve fibers, and several subtypes of neuronal populations residing in the myenteric plexus. Comparison with existing methodsCompared with existing procedures for whole-mount preparations, the proposed method achieves a quicker and more efficient preparation of high-quality LMMPs from intestinal segments in sufficient quantity. ConclusionsThis work provides a rapid method for efficiently preparing whole mounts of the intestines. Our method can be used for in situ observation of the morphological and functional alterations of the myenteric plexus for further studies on the ENS.

Characterisation of calcitonin gene-related peptide-immunoreactive neurons in the myenteric plexus of rat colon

A mechanical or chemical stimulus applied to the intestinal mucosa induces motility reflexes in the rat colon. Enteric neurons containing calcitonin gene-related peptide (CGRP) have been suggested as intrinsic primary afferent neurons responsible for mediating such reflexes. In the present study, immunohistochemistry was performed on whole-mount stretch preparations to investigate chemical profiles, morphological characteristics and projections of CGRP-containing neurons in the myenteric plexus of the rat colon. CGRP-positive neuronal cell bodies were detected in preparations incubated with colchicine-containing medium, whereas CGRP-positive nerve fibres were found in colchicine-untreated preparations. These neurons had large oval or round cell bodies that were also immunoreactive for the calcium-binding protein calretinin and neurofilament 200. Myenteric neurons positive for both calretinin and neurofilament 200 had several long processes that emerged from the cell body, consistent with Dogiel type II morphology. Application of the neural tracer DiI to the intestinal mucosa revealed that DiI-labelled myenteric neurons each had an oval or round cell body immunoreactive for calretinin. Thus, CGRP-containing myenteric neurons are Dogiel type II neurons and are immunoreactive for calretinin and neurofilament 200 in the rat colon. These neurons probably project to the intestinal mucosa.